Transport of anions in isolated barley vacuoles : I. Permeability to anions and evidence for a cl-uptake system

The anion contents of young barley leaves and of mesophyll protoplasts from the leaves was compared. Anion loss from the protoplasts during isolation was small. Although only about 60% of the leaf cells were mesophyll cells, phosphate and sulfate contents of the mesophyll cells accounted for almost 90% of the leaf contents. Chloride accumulated in the leaf epidermis. The rapid isolation of vacuoles from mesophyll protoplasts permitted the determination of vacuolar ion concentrations. Sodium and nitrate levels were very low in the cytoplasm, and much higher in the vacuole. When barley plants were grown in the presence of low NaCl levels, chloride concentrations were comparable in cytoplasm and vacuole, and similar observations were made with sulfate. Cytoplasmic phosphate concentrations were close to 30 millimolar and potassium concentrations 100 millimolar. During a 30 minute incubation period at room temperature, anion contents of isolated vacuoles decreased considerably. Efflux of NO₃⁻ was faster than that of Cl⁻. Phosphate and sulfate crossed the tonoplast only slowly. 4,4′-Diisothiocyano-2,2′-stilbenedisulfonic acid partially inhibited the efflux of nitrate and, to a lesser extent, that of chloride. Decreased efflux was also observed in the presence of MgATP. In remarkable contrast, p-chloromercuribenzene sulfonate and HgCl₂ stimulated the efflux of nitrate and chloride, but not of phosphate. Labeled chloride was taken up by isolated vacuoles. The apparent K_m for chloride uptake at low chloride concentrations was 2.3 millimolar. At elevated chloride concentrations, chloride did not display saturation characteristics but, rather, characteristics of a diffusional process. Uptake was stimulated by ATP.     

霍山石斛组培苗移栽后微观结构与微区元素的变化研究

采用扫描电镜及X射线能谱仪技术,研究霍山石斛组培苗移栽后各器官组织的微观结构和所含元素的变化,以了解霍山石斛组培苗生长发育以及药效成分的状况。结果表明:(1)霍山石斛组培苗移栽2个月后,根增粗近1倍,具有根被,且根被细胞壁的网络状增厚更加明显,表皮、皮层、中柱发育分化更加完善,Mg、Si、Cl、S、K、Ca元素含量提高,其中Ca增长了2.76倍。(2)茎表面纵向凹陷增多,内部结构致密,维管束发育较为完善,细胞内充满物质,Mg、Si、Cl、S、K、Ca、Fe、Cu、Zn元素含量增多,尤其是K和Fe分别增长了3.25倍和4.61倍。(3)叶增厚,具有了角质层,气孔形状更加饱满,叶肉细胞内含物丰富,Mg、Si、Cl、S、K、Ca、Fe、Mn、Zn的含量显著上升,其中K升高了17倍。(4)与根、叶相比,茎中所富集的元素种类最多。研究认为,霍山石斛试管苗移栽后,生长发育趋向完善,各项功能增强,细胞内含物质增多,元素成分丰富并且含量提高,体现出对移栽环境的良好适应性。     

Infection Process of Botrytis cinerea on Eucalypt Leaves

This study aimed to elucidate the infection process of Botrytis cinerea on eucalypt leaves. Tests were conducted to evaluate the influence of leaf side (adaxial or abaxial), leaf age and luminosity on conidial germination, appressorium formation and grey mould (GM) severity. The adaxial and abaxial surfaces of detached eucalypt leaves were inoculated with a conidial suspension of B. cinerea and kept under constant light or dark. Subsequently, the adaxial surface of young and old leaves was inoculated and kept in the dark. To evaluate the percentage of conidia germination and appressorium formation, leaf samples were collected 6 hours after inoculation (hai), clarified (alcohol and chloral hydrate) and evaluated under a light microscope. The severity of GM was assessed 10 days after inoculation. For scanning electron microscopy analysis, samples were collected from 2 to 168 hai. A higher percentage of conidia germination (92%) and GM severity (21%) occurred on the adaxial surfaces of leaves kept in the dark. There was no statistical difference between the surfaces of young and old leaves for conidia germination. No appressorium was formed by B. cinerea. The GM severity on young leaves (17.3%) was 34 times higher than on old leaves (0.5%). The micrographs showed germinating conidia emitting 1–4 germ tubes in samples at 4 hai. The fungus penetration occurred through intact leaf surfaces, and both extra‐ and intracellular colonization of the mesophyll cells by the hyphae of the pathogen were observed at 120 hai. Sporulation occurred on the adaxial and abaxial surfaces (macronematous conidiophores) and below the epidermis (micronematous conidiophores).     

不同竹龄雷竹中硅及其他营养元素吸收和积累特征

在浙江省临安市的雷竹主产区,分别采集不同竹龄(1～4 a)和不同器官(叶、枝、秆)的雷竹样品,分析了Si和其他营养元素含量、吸收和积累特征,以及Si和其他营养元素之间的相互关系.结果表明： 雷竹各器官中C含量的大小顺序为竹秆>竹枝>竹叶,Si、N、P、K、Ca、Mg、Al、Fe和Mn含量的大小顺序为竹叶>竹枝>竹秆.除Mn主要积累在竹叶中外,其他9种营养元素主要积累在1年生雷竹的秆中.3～4年生雷竹竹叶的Si平均含量为13.66 g · kg^-1. 雷竹属于Si积累植物.随竹龄的增加,雷竹叶中的N、P、K和Mg含量减少,C、Al和Mn含量增加.雷竹对Si的吸收主要集中在第2年(57.1%),对N和K的吸收主要集中在前两年(67.7%～93.7%),此后N和K从植株体内流出,其流失量分别占总积累量的19.1%～39.1%.雷竹中Si与Ca、Al、Mn呈显著正相关,与N、P、K、Mg呈显著负相关.     

Distribution and mobility of manganese in the hyperaccumulator plant Phytolacca acinosa Roxb. (Phytolaccaceae)

The distribution and mobility of manganese (Mn) in the hyperaccumulator plant species Phytolacca acinosa Roxb. (Phytolaccaceae) were investigated in a hydroponic system. The plants were exposed to 2 or 5 mM Mn for up to 28 days. For any given plant, the Mn content in the mature leaves (nos. 5–9) was always higher than that in the old (nos. 1–4) and young leaves (nos. 10–14). Within the different parts of a leaf, Mn was preferentially accumulated in the leaf marginal area, where the observed level was threefold higher than that in the midrib. Cross-sectional analysis of the leaf revealed that the concentration of Mn was higher in the leaf epidermis than in the mesophyll. Cell fractionation analysis with P. acinosa leaves showed that most of the Mn (78.4%) was present in the final supernatant fraction (following centrifugation at 20,000 g for 45 min). The distribution of Mn in the leaves of P. acinosa was controlled mainly by the transpiration rate. Our investigation demonstrated that Mn was readily transported from the roots to shoots of P. acinosa but that it could not be remobilized readily after it reached leaves.     

Distribution and chemical speciation of aluminum in the Al accumulator plant,Melastoma malabathricum L.

The Al accumulation mechanisms in an Al accumulator plant, Melastoma malabathricum L. (Melastoma), was investigated. Al was located in the upper epidermal cells and also distributed in mesophyll cells in leaf sections. In root sections, Al was found in all the root tissues, particularly in the epidermis and endodermis. Al concentrations in young leaves, mature leaves, old leaves, and roots were 8.0, 9.2, 14.4, and 10.1 mg g¹, respectively. Approximately 45% of total Al in oldest leaves, and approximately 60% of total Al in leaves of other positions and roots were extracted in Tris-HCl buffer (pH 7.0). Since Al in the residual parts was mostly dissolved in hot 0.5 M H₂SO₄ containing 2% cetyl trimethylammonium bromide, residual Al seemed to consist mainly of monomeric Al and Al bound to pectic substances and hemicellulose. Al in the Tris-HCl extract consisted of non-monomeric Al (complexed form). Oxalate concentration in the Tris-HCl extract in leaves was significantly higher in the +Al treatment than in the –Al treatment and there was a positive correlation between the Al concentration and oxalate concentration. ²⁷Al NMR spectrum of fresh leaves indicated the presence in the order of monomeric Al, Al-oxalate, Al-(oxalate)₂, and Al-(oxalate)₃ in intact leaves.     

Cellular compartmentation of cadmium and zinc in relation to other elements in the hyperaccumulator Arabidopsis halleri

The cellular compartmentation of elements was analysed in the Zn hyperaccumulator Arabidopsis halleri (L.) O'Kane & Al-Shehbaz (=Cardaminopsis halleri) using energy-dispersive X-ray microanalysis of frozen-hydrated tissues. Quantitative data were obtained using oxygen as an internal standard in the analyses of vacuoles, whereas a peak/background ratio method was used for quantification of elements in pollen and dehydrated trichomes. Arabidopsis halleri was found to hyperaccumulate not only Zn but also Cd in the shoot biomass. While large concentrations of Zn and Cd were found in the leaves and roots, flowers contained very little. In roots grown hydroponically, Zn and Cd accumulated in the cell wall of the rhizodermis (root epidermis), mainly due to precipitation of Zn/Cd phosphates. In leaves, the trichomes had by far the largest concentrations of Zn and Cd. Inside the trichomes there was a striking sub-cellular compartmentation, with almost all the Zn and Cd being accumulated in a narrow ring in the trichome base. This distribution pattern was very different from that for Ca and P. The epidermal cells other than trichomes were very small and contained lower concentrations of Zn and Cd than mesophyll cells. In particular, the concentrations of Cd and Zn in the mesophyll cells increased markedly in response to increasing Zn and Cd concentrations in the nutrient solution. This indicates that the mesophyll cells in the leaves of A. halleri are the major storage site for Zn and Cd, and play an important role in their hyperaccumulation. Received: 4 April 2000 / Accepted: 16 May 2000     

四种金花茶组植物叶片金属元素含量及富集特性研究

以四种金花茶组植物为研究对象,采用原子吸收光谱法和原子荧光法,测定其嫩叶、老叶及对应土壤中Mg、Ca、Mn、Fe、Zn、Ni、Se、Pb、Cd、Hg、As共11种元素的含量,并分别计算嫩叶和老叶对土壤金属元素的富集系数.结果表明:(1)4种金花茶组植物叶片富含Mg、Ga、Mn、Fe、Zn、Ni等营养元素,各元素在叶片中含量为Ca>Mg>Mn>Fe>Zn>Ni>Se;Pb、Cd、As、Hg等重金属元素含量较低,均达到无公害茶叶标准.(2)老叶和嫩叶中各金属元素含量差异较大,老叶中的Ca、Mn、Fe、Zn、Pb、Cd、Hg、As、Se元素含量均大于嫩叶,尤以Ca、Mn、Fe差异显著;嫩叶中的Mg和Ni含量大于老叶.(3)金花茶组植物对不同金属元素的富集能力不同,对各元素富集能力强弱为Ca、Mn、Mg>Zn、Ni、Hg>Pb、Se>Fe、As,老叶和嫩叶的富集规律存在差异.(4)不同金花茶组植物对金属元素的富集能力有较大差异,龙州金花茶(Camellia longzhouensis)和黄花抱茎茶(C.murauchii)对Mg、Ca、Mn、Zn、Ni、Se、Pb的富集能力均大于金花茶(C.nitidissima)和毛籽金花茶(C.ptilosperma).其中,龙州金花茶对Mg、Mn、Se的富集能力最强,黄花抱茎茶对Ca、Pb、Hg富集能力最强,金花茶对Hg的富集能力较强,对其它元素的富集能力均较弱;毛籽金花茶对Ca、Mn、Ni、Zn的富集能力均最弱.该研究结果为金花茶组植物的进一步开发和利用提供了理论依据.     

Compartmentation and transport of zinc in barley primary leaves as basic mechanisms involved in zinc tolerance

The heavy metal zinc was administered to barley seedlings by increasing its concentration in the hydroponic medium. The most dramatic effect was a severe inhibition of root elongation with little effect on root biomass production. The growth of primary leaves was little affected although the zinc content of the primary leaves increased several-fold. A detailed compartment analysis was performed for 10-d-old barley primary leaves. Under low zinc nutrition (2mmol m ⁻³), highest zinc contents were observed in the cytoplasm of mesophyll protoplasts. At inhibitory zinc concentrations in the hydroponic medium (400 μmol m ⁻³), zinc levels dramatically and preferentially increased in the apoplastic space. Elevated zinc levels were also observed in the epidermal cells, and to a lesser extent, in mesophyll vacuoles. The cytoplasmic content of mesophyll protoplasts was unchanged, indicating perfect zinc homeostasis within the leaf. In order to understand the transport mechanisms underlying the steady-state distribution profile, we used ⁶⁵Zn to conduct uptake experiments with leaves whose lower epidermis had been stripped. The leaves were placed on zinc solutions of varying concentrations containing ⁶⁵Zn for 5 min to 6 h. After the incubation, the leaves were fractionated into mesophyll and epidermis protoplasts and residue, the latter mainly representing cell wall. Adsorption of Zn to the extracellular matrix was 100 times faster than Zn uptake into the cells. By far the largest portion taken up into the mesophyll protoplasts rapidly appeared in the vacuolar compartment. These results demonstrate the importance of compartmentation and transport as homeostatic mechanisms within the leaves to handle high, possibly toxic, zinc levels in the shoot.     

Responses of the Populus × euramericana clone I-214 to excess zinc: Carbon assimilation,structural modifications,metal distribution and cellular localization

Poplar (Populus), the model system in tree research, is a fast-growing and high biomass plant which is promising for energy, paper and pulp production, and for growth in soils contaminated with metals. Contamination of soils and water with heavy metals has become a widespread problem; environmental pollution by excess zinc (Zn), one of the more important contaminants, occurs frequently and yet the responses of Populus to high Zn concentrations are still not clearly understood.We investigated the effects of Zn on the functional and structural parameters in the Populus × euramericana clone I-214 by Zn localization in frozen-hydrated leaves and roots by cryo-scanning electron microscopy (cryo-SEM)/energy-dispersive X-ray microanalysis (EDXMA). The experiment was conducted on cuttings grown in nutrient solutions with an increasing Zn concentration gradient (0.001–10 mM).Biomass partitioning and Zn uptake were affected by the metal treatments, showing organ- and tissue-dependent responses. In particular, Zn accumulated in old leaves and moved from shoot to root as the Zn concentration in the growth medium increased. At the highest treatment concentration (10 mM), Zn was preferentially localized in photosynthetic tissues of shoots, and in epidermis and cortex tissues of roots. Gas exchange and chlorophyll measurements showed impairments in leaf biochemistry rather than in stomatal function. Modifications in foliage area, stomatal density and leaf layer thickness were investigated to reduce and/or compensate the negative effects of excess Zn on CO₂ assimilation.To counteract Zn toxicity, clone I-214 adopted different defense/tolerance mechanisms involving complex structural, physiological and biochemical processes, attributed to both Zn excluders and accumulators. This study demonstrates the advantages of combining cryo-SEM/EDXMA, gas exchange and chemical analyses for studying metal localization and structural as well as physiological responses in plants.     

Rare earth elements,aluminium and silicon distribution in the fern Dicranopteris linearis revealed by μPIXE Maia analysis

BackgroundThe fern Dicranopteris linearis is a hyperaccumulator of rare earth elements (REEs), aluminium (Al) and silicon (Si). However, the physiological mechanisms of tissue-level tolerance of high concentrations of REE and Al, and possible interactions with Si, are currently incompletely known.MethodsA particle-induced X-ray emission (μPIXE) microprobe with the Maia detector, scanning electron microscopy with energy-dispersive spectroscopy and chemical speciation modelling were used to decipher the localization and biochemistry of REEs, Al and Si in D. linearis during uptake, translocation and sequestration processes.ResultsIn the roots >80 % of REEs and Al were in apoplastic fractions, among which the REEs were most significantly co-localized with Si and phosphorus (P) in the epidermis. In the xylem sap, REEs were nearly 100 % present as REEH₃SiO₄²⁺, without significant differences between the REEs, while 24–45 % of Al was present as Al-citrate and only 1.7–16 % Al was present as AlH₃SiO₄²⁺. In the pinnules, REEs were mainly concentrated in necrotic lesions and in the epidermis, and REEs and Al were possibly co-deposited within phytoliths (SiO₂). Different REEs had similar spatial localizations in the epidermis and exodermis of roots, the necrosis, veins and epidermis of pinnae of D. linearis.ConclusionsWe posit that Si plays a critical role in REE and Al tolerance within the root apoplast, transport within the vascular bundle and sequestration within the blade of D. linearis.     

Age-Dependent Variations of Volatile Emissions and Inhibitory Activity Toward <Emphasis Type="Italic">Botrytis cinerea</Emphasis> and <Emphasis Type="Italic">Fusarium oxysporum</Emphasis> in Tomato Leaves Treated with Chitosan Oligosaccharide

We investigated variations in the level and composition of volatiles emitted by tomato leaves at different ages. Our focus also included their antifungal properties and responses to chitosan oligosaccharide. Based on leaf position, the release of volatiles decreased over time. Young leaves produced high levels of C6-aldehyde, which is mainly composed of hexenal, while the volatiles emitted by more mature leaves largely comprised terpenes, particularly β-phellandrene and caryophyllane. In young upper leaves, the main components (up to 86% of the total) were hexenal, β-phellandrene, and caryophyllane. Their levels decreased steadily over time, from 386.3 μg g⁻¹ fresh weight (FW) in young leaves to 113.2 μg g⁻¹ FW in old tissues. Volatiles emitted from young leaves exhibited the best antifungal activity against spore germination and hyphal growth by Botrytis cinerea and Fusarium oxysporum. Leaves became more susceptible to oligosaccharide treatment with increasing age. When young tissues were exposed to chitosan, we found declines in both the quantity of volatiles and their ability to inhibit fungal growth. Compared with the control, the amount of volatiles from young tissues was 88.4% lower after such treatment. In contrast, contents of volatiles from old and adult leaves were dramatically increased by chitosan oligosaccharide. Likewise, their inhibitory effect was significantly enhanced. Therefore, our results suggest that these volatiles are responsible for antifungal activity and may play a role in age-related resistance by tomato.     

Elemental composition of the cyanobacterium Anabaena flos-aquae collected from different depths within a stratified lake

The elemental composition of cells of Anabaena flos-aquae was investigated by energy-dispersive electron probe X-ray microanalysis (10?kV) of mixed phytoplankton samples obtained from different depths within a stratified eutrophic freshwater lake. Preliminary studies had indicated that at 10?kV, X-ray information was derived mainly from the peripheral region (up to 1–2?μm) of cells. Routinely detectable elements (present in at least 50% of cells) included Mg (overall mean 112?mmol?kg^?1 dry mass), Si (1850), P(313), S (113), Cl (92), K (300) and Ca (56). Peaks of Na and Al were also frequently present. Individual elements showed a wide range of intracellular concentrations at each depth within the lake. Si had a clear bimodal distribution at depths of 1–8?m, indicating both low-Si (mean concentrations 54–290–3800?mmol?Si?kg^?1) and high-Si (2770–3800?mmol?Si?kg^?1) cell subpopulations. Low-Si cells had significantly higher concentrations of other detectable elements compared with high-Si cells, but could not be distinguished from the latter in terms of morphology or stage of cell cycle (comparison of dividing and non-dividing cells). High-Si cells were intermixed with low-Si cells in individual Anabaena colonies and decreased proportionally with depth, ranging from approximately 75% in the surface sample to 10% at 8?m. With the exception of Si, mean elemental concentrations of Anabaena populations were closely similar throughout the epilimnion. These differed from the metalimnion sample, which had significantly higher concentrations of Mg, P, S and K, and a lower concentration of Si. Mean ratios of certain elements (Mg, P, S and K) and ion groups (monovalent/divalent cations) were highly constant throughout the sampled water column. At each depth, most of the cell elements were significantly and positively inter-correlated (Pearson analysis), with Mg, P, S and K typically constituting the major group (factor analysis). Correlations with Si were invariably negative, suggesting a separate (possibly cell wall) location of this element in the cell.     

Site of Monoterpene Biosynthesis in Majorana hortensis Leaves

Excised epidermis of Majorana hortensis Moench (sweet marjoram) leaves incorporates label from [U-¹⁴C]sucrose into monoterpenes as efficiently as do leaf discs, while mesophyll tissue has only a very limited capacity to synthesize monoterpenes from exogenous sucrose. These results strongly suggest that epidermal cells, presumably the epidermal oil glands, are the primary site of monoterpene biosynthesis in marjoram. Using a leaf disc assay, it was demonstrated that label from [U-¹⁴C]sucrose is incorporated into monoterpenes most efficiently in very young leaves.     

Exogenous proline effects on water relations and ions contents in leaves and roots of young olive

The ability of exogenous compatible solutes, such as proline, to counteract salt inhibitory effects was investigated in 2-year-old olive trees (Olea europaea L. cv. Chemlali) subjected to different saline water irrigation levels supplied or not with exogenous proline. Leaf water relations [relative water content (RWC), water potential], photosynthetic activity, leaf chlorophyll content, and starch contents were measured in young and old leaves. Salt ions (Na⁺, K⁺, and Ca²⁺), proline and soluble sugars contents were determined in leaf and root tissues. Supplementary proline significantly mitigated the adverse effects of salinity via the improvement of photosynthetic activity (Pn), RWC, chlorophyll and carotenoid, and starch contents. Pn of young leaves in the presence of 25 mM proline was at 1.18 and 1.38 times higher than the values recorded under moderate (SS1) and high salinity (SS2) treatments, respectively. Further, the proline supply seems to have a more important relaxing effect on the photosynthetic chain in young than in old leaves of salt-stressed olive plants. The differential pattern of proline content between young and old leaves suggests that there would be a difference between these tissues in distinguishing between the proline taken from the growing media and that produced as a result of salinity stress. Besides, the large reduction in Na⁺ accumulation in leaves and roots in the presence of proline could be due to its interference in osmotic adjustment process and/or its dilution by proline supply. Moreover, the lower accumulation of Na⁺ in proline-treated plants, compared to their corresponding salinity treatment, displayed the improved effect of proline on the ability of roots to exclude the salt ions from the xylem sap flowing to the shoot, and thus better growth rates.     

Tolerance,accumulation and distribution of zinc and cadmium in hyperaccumulator Potentilla griffithii

In this study, zinc (Zn) and cadmium (Cd) tolerance, accumulation and distribution was conducted in Potentilla griffithii H., which has been identified as a new Zn hyperaccumulator found in China. Plants were grown hydroponically with different levels of Zn²⁺ (20, 40, 80 and 160 mg L^?1) and Cd²⁺ (5, 10, 20 and 40 mg L^?1) for 60 days. All plants grew healthy and attained more biomass than the control, except 40 mg L^?1 Cd treatment. Zn or Cd concentration in plants increased steadily with the increasing addition of Zn or Cd in solution. The maximum metal concentrations in roots, petioles and leaves were 14,060, 19,600 and 11,400 mg kg^?1 Zn dry weight (DW) at 160 mg L^?1 Zn treatment, and 9098, 3077 and 852 mg kg^?1 Cd DW at 40 mg L^?1 Cd treatment, respectively. These results suggest that P. griffithii has a high ability to tolerate and accumulate Cd and Zn, and it can be considered not only as Zn but also as a potential cadmium hyperaccumulator. Light microscope (LM) with histochemical method, scanning electron microscope combined with energy dispersive spectrometry (SEM-EDS) and transmission electron microscope (TEM) were used to determine the distribution of Zn and Cd in P. griffithii at tissue and cellular levels. In roots, SEM-EDS confirmed that the highest Zn concentration was found in xylem parenchyma cells and epidermal cells, while for Cd, a gradient was observed with the highest Cd concentration in rhizodermal and cortex cells, followed by central cylinder. LM results showed that Zn and Cd distributed mainly along the walls of epidermis, cortex, endodermis and some xylem parenchyma. In leaves, Zn and Cd shared the similar distribution pattern, and both were mostly accumulated in epidermis and bundle sheath. However, in leaves of 40 mg L^?1 Cd treatment, which caused the phytotoxicity, Cd was also found in the mesophyll cells. The major storage site for Zn and Cd in leaves of P. griffithii was vacuoles, to a lesser extent cell wall or cytosol. The present study demonstrates that the predominant sequestration of Zn and Cd in cell walls of roots and in vacuoles of epidermis and bundle sheath of leaves may play a major role in strong tolerance and hyperaccumulation of Zn and Cd in P. griffithii.     

Major Element Composition of Epidermal and Mesophyll Tissues2Commelina Communis L. and Vicia Faba L.: Some Further Considerations of the Role of Ions in Stomatal Functioning

Epidermal and mesophyll tissues of Commelina communis L. andVicia faba L. were analysed by atomic absorption spectrometryfor the major plant inorganic cations and anions (K, Na, Ca,Mg, P, NO₃-N, Cl) when stomata of the leaf were open and closed.Water-soluble and residual levels of the elements were estimatedand a charge balance of the soluble fraction made. The major portion of K, Na, Cl, and P was extracted in the water-solublefraction of the epidermal and mesophyll tissues of both species.In both species the bulk of Ca remained in the insoluble residueof the epidermis whereas in mesophyll tissue it was equallydistributed be-between the two fractions in C. communis butmainly in the insoluble residue in V. faba. Magnesium was predominantlyfound in the water-soluble fraction of V. faba mesophyll tissueand distributed approximately equally between the two fractionsin the epidermal tissue. In C. communis Mg was slightly moreabundant in the water-soluble fraction of both mesophyll andepidermis. In both species no statistically significant differences inthe levels of the elements could be detected between epidermaland mesophyll tissues from leaves with open stomata and thesame tissues from leaves with closed stomata, suggesting thatthere was no major flux of ions between mesophyll and epidermisduring stomatal movements. Regardless of whether the stomata were open or closed, therewere considerably more water-soluble inorganic cations thananions present in all tissues of both species with K being themajor cation and Cl being the major anion. In V.faba and C-communis epidermis there was 49–53 per cent and 56%68per cent excess cation respectively. In the mesophyll tissuethe excess cation was 63–75 per cent and 75%78 per centin V.faba and C. communis respectively. When the partitioning of the levels of the elements betweenepidermis and mesophyll of a leaf is considered, except forNO₃-N in both species and Na in V. faba, 20 per cent or lessof each element was present in the epidermis.     

Different patterns of ecosystem carbon accumulation between a young and an old-growth subtropical forest in Southern China

Using long-term (22 years) measurements from a young and an old-growth subtropical forest in southern China, we found that both forests accumulated carbon from 1982 to 2004, with the mean carbon accumulation rate at 227 ± 59 g C m⁻² year⁻¹ for young forest and 115 ± 89 g C m⁻² year⁻¹ for the old-growth forest. Allocation of the accumulated carbon was quite different between these two forests: the young forest accumulated a significant amount of carbon in plant live biomass, whereas the old-growth forest accumulated a significant amount of carbon in the soil. From 1982 to 2004, net primary productivity (NPP) increased for the young forest, and did not change significantly for the old-growth forest. The increase in NPP of the young forest resulted from recruitment of some dominant tree species characteristic of the subtropical mature forest in the region and an increase in tree density; decline of NPP of the old-growth forest was caused by increased mortality of the dominant trees.     

The role of the epidermis in the generation of the circadian rhythm of carbon dioxide fixation in leaves of Bryophyllum fedtschenkoi

The role of the epidermis in the generation of the endogenous circadian rhythm of CO₂ exchange in leaves of Bryophyllum fedtschenkoi has been examined. At 25° C the rhythm of CO₂ output exhibited by whole leaves kept in continuous darkness and an initially CO₂-free air stream also occurs in isolated pieces of mesophyll. The sensitivity to light of the rhythms in whole leaves and in isolated mesophyll appears to be identical. At 15° C, however, no rhythm is observed in isolated mesophyll tissue, despite there being a conspicuous rhythm in intact leaves. The rhythm of net CO₂ assimilation in whole leaves kept in continuous light and a stream of normal air at either 25° C or at 15° C is abolished by removal of the epidermis, although at 15° C and under the higher of the two light levels used, there is an indication that rhythmicity may begin to reappear after the third day of the experiment. Thus, only under certain environmental conditions is the rhythm of CO₂ exchange in Bryophyllum leaves independent of the epidermis. The results indicate that the rhythm of carbon dioxide fixation in continuous darkness and CO₂-free air is generated primarily in the mesophyll cells, whereas the rhythm in continuous light and normal air is generated in the stomatal guard cells or in an interaction of these cells with the mesophyll cells.Abbreviation PEPCase phosphoenolpyruvate carboxylase